Flow cytometric pattern (intracellular staining) of HeLa cells (Human epithelial cell line from cervix adenocarcinoma cell suspension) stained with ab668  (0.6 µg/ml concentration)

Immunohistochemistry (paraffin-embedded sections) analysis of human kidney tissue labelling cytokeratin-18 with ab668 at 10µg/ml concentration.

Flow cytometric analysis (intracellular staining) of HeLa cells (human epithelial cell line from cervix adenocarcinoma cell suspension) stained with 0.6 µg/ml ab668 (in red), and unstained HeLa cells (black).

Overlay histogram showing HCT 116 (Human colorectal carcinoma cell line) cells stained with ab668 (red line).

The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Triton for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab668, 1µg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HCT 116 cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Triton used under the same conditions.

ab668 staining Cytokeratin 18 in cat lung tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in citric acid. Samples were incubated with primary antibody (1/500 in TBS/BSA/azide) for 2 hours at 21°C. A biotin-conjugated goat anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.

Coloured arrowheads indicate positivity.
Good immunolabeling of bronchial tree epithelia (green), alveolar lining epithelium (red ).
Goblet cells are negative (asterisk).
Heavily stained structures are submucosal mucous glands.

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ab668 staining cytokeratin 18 in mouse epithelium tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

Tissue was fixed with formaldehyde and blocked with 1% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/500 in TBS/BSA/azide) for 2 hours at 21°C. A biotin-conjugated Goat anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.

Composite image of d14 embryo shows developing epithelia.

Upper image: positive simple epithelium of renal tubules

Lower image: negative stratified epithelium of epidermis

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